In Situ Encapsulation of SnS2/MoS2 Heterojunctions by Amphiphilic Graphene for High-Energy and Ultrastable Lithium-Ion Anodes.

Lithium-ion batteries with transition metal sulfides (TMSs) anodes promise a high capacity, abundant resources, and environmental friendliness, yet they suffer from fast degradation and low Coulombic efficiency. Here, a heterostructured bimetallic TMS anode is fabricated by in situ encapsulating SnS2/MoS2 nanoparticles within an amphiphilic hollow double-graphene sheet (DGS). The hierarchically porous DGS consists of inner hydrophilic graphene and outer hydrophobic graphene, which can accelerate electron/ion migration and strongly hold the integrity of alloy microparticles during expansion and/or shrinkage. Moreover, catalytic Mo converted from lithiated MoS2 can promote the reaction kinetics and suppress heterointerface passivation by forming a building-in-electric field, thereby enhancing the reversible conversion of Sn to SnS2. Consequently, the SnS2/MoS2/DGS anode with high gravimetric and high volumetric capacities achieves 200 cycles with a high initial Coulombic efficiency of >90%, as well as excellent low-temperature performance. When the commercial Li(Ni0.8Co0.1Mn0.1)O2 (NCM811) cathode is paired with the prelithiated SnS2/MoS2/DGS anode, the full cells deliver high gravimetric and volumetric energy densities of 577 Wh kg-1 and 853 Wh L-1, respectively. This work highlights the significance of integrating spatial confinement and atomic heterointerface engineering to solve the shortcomings of conversion-/alloying typed TMS-based anodes to construct outstanding high-energy LIBs.

GPCR-G protein selectivity revealed by structural pharmacology.

Receptor-G protein promiscuity is frequently observed in class A G protein-coupled receptors (GPCRs). In particular, GPCRs can couple with G proteins from different families (Gαs, Gαq/11, Gαi/o, and Gα12/13) or the same family subtypes. The molecular basis underlying the selectivity/promiscuity is not fully revealed. We recently reported the structures of kappa opioid receptor (KOR) in complex with the Gi/o family subtypes [Gαi1, GαoA, Gαz, and Gustducin (Gαg)] determined by cryo-electron microscopy (cryo-EM). The structural analysis, in combination with pharmacological studies, provides insights into Gi/o subtype selectivity. Given the conserved sequence identity and activation mechanism between different G protein families, the findings within Gi/o subtypes could be likely extended to other families. Understanding the KOR-Gi/o or GPCR-G protein selectivity will facilitate the development of more precise therapeutics targeting a specific G protein subtype.

Design and structural validation of peptide-drug conjugate ligands of the kappa-opioid receptor.

Despite the increasing number of GPCR structures and recent advances in peptide design, the development of efficient technologies allowing rational design of high-affinity peptide ligands for single GPCRs remains an unmet challenge. Here, we develop a computational approach for designing conjugates of lariat-shaped macrocyclized peptides and a small molecule opioid ligand. We demonstrate its feasibility by discovering chemical scaffolds for the kappa-opioid receptor (KOR) with desired pharmacological activities. The designed De Novo Cyclic Peptide (DNCP)-ß-naloxamine (NalA) exhibit in vitro potent mixed KOR agonism/mu-opioid receptor (MOR) antagonism, nanomolar binding affinity, selectivity, and efficacy bias at KOR. Proof-of-concept in vivo efficacy studies demonstrate that DNCP-ß-NalA(1) induces a potent KOR-mediated antinociception in male mice. The high-resolution cryo-EM structure (2.6 Å) of the DNCP-ß-NalA-KOR-Gi1 complex and molecular dynamics simulations are harnessed to validate the computational design model. This reveals a network of residues in ECL2/3 and TM6/7 controlling the intrinsic efficacy of KOR. In general, our computational de novo platform overcomes extensive lead optimization encountered in ultra-large library docking and virtual small molecule screening campaigns and offers innovation for GPCR ligand discovery. This may drive the development of next-generation therapeutics for medical applications such as pain conditions.

OsIPK2, a Rice Inositol Polyphosphate Kinase Gene, Is Involved in Phosphate Homeostasis and Root Development.

Phosphorus (P) is a growth-limiting nutrient for plants, which is taken up by root tissue from the environment as inorganic phosphate (Pi). To maintain an appropriate status of cellular Pi, plants have developed sophisticated strategies to sense the Pi level and modulate their root system architecture (RSA) under the ever-changing growth conditions. However, the molecular basis underlying the mechanism remains elusive. Inositol polyphosphate kinase (IPK2) is a key enzyme in the inositol phosphate metabolism pathway, which catalyzes the phosphorylation of IP3 into IP5 by consuming ATP. In this study, the functions of a rice inositol polyphosphate kinase gene (OsIPK2) in plant Pi homeostasis and thus physiological response to Pi signal were characterized. As a biosynthetic gene for phytic acid in rice, overexpression of OsIPK2 led to distinct changes in inositol polyphosphate profiles and an excessive accumulation of Pi levels in transgenic rice under Pi-sufficient conditions. The inhibitory effects of OsIPK2 on root growth were alleviated by Pi-deficient treatment compared with wild-type plants, suggesting the involvement of OsIPK2 in the Pi-regulated reconstruction of RSA. In OsIPK2-overexpressing plants, the altered acid phosphatase (APase) activities and misregulation of Pi-starvation-induced (PSI) genes were observed in roots under different Pi supply conditions. Notably, the expression of OsIPK2 also altered the Pi homeostasis and RSA in transgenic Arabidopsis. Taken together, our findings demonstrate that OsIPK2 plays an important role in Pi homeostasis and RSA adjustment in response to different environmental Pi levels in plants.

Ligand and G-protein selectivity in the κ-opioid receptor.

The κ-opioid receptor (KOR) represents a highly desirable therapeutic target for treating not only pain but also addiction and affective disorders1. However, the development of KOR analgesics has been hindered by the associated hallucinogenic side effects2. The initiation of KOR signalling requires the Gi/o-family proteins including the conventional (Gi1, Gi2, Gi3, GoA and GoB) and nonconventional (Gz and Gg) subtypes. How hallucinogens exert their actions through KOR and how KOR determines G-protein subtype selectivity are not well understood. Here we determined the active-state structures of KOR in a complex with multiple G-protein heterotrimers-Gi1, GoA, Gz and Gg-using cryo-electron microscopy. The KOR-G-protein complexes are bound to hallucinogenic salvinorins or highly selective KOR agonists. Comparisons of these structures reveal molecular determinants critical for KOR-G-protein interactions as well as key elements governing Gi/o-family subtype selectivity and KOR ligand selectivity. Furthermore, the four G-protein subtypes display an intrinsically different binding affinity and allosteric activity on agonist binding at KOR. These results provide insights into the actions of opioids and G-protein-coupling specificity at KOR and establish a foundation to examine the therapeutic potential of pathway-selective agonists of KOR.

Conformational cycle of human polyamine transporter ATP13A2.

Dysregulation of polyamine homeostasis strongly associates with human diseases. ATP13A2, which is mutated in juvenile-onset Parkinson's disease and autosomal recessive spastic paraplegia 78, is a transporter with a critical role in balancing the polyamine concentration between the lysosome and the cytosol. Here, to better understand human ATP13A2-mediated polyamine transport, we use single-particle cryo-electron microscopy to solve high-resolution structures of human ATP13A2 in six intermediate states, including the putative E2 structure for the P5 subfamily of the P-type ATPases. These structures comprise a nearly complete conformational cycle spanning the polyamine transport process and capture multiple substrate binding sites distributed along the transmembrane regions, suggesting a potential polyamine transport pathway. Integration of high-resolution structures, biochemical assays, and molecular dynamics simulations allows us to obtain a better understanding of the structural basis of how hATP13A2 transports polyamines, providing a mechanistic framework for ATP13A2-related diseases.

Investment of needle nitrogen to photosynthesis controls the nonlinear productivity response of young Chinese fir trees to nitrogen deposition.

Plant carbon (C) assimilation is expected to nonlinearly increase with continuously increasing nitrogen (N) deposition, causing a N saturation threshold for productivity. However, the response of plant productivity to N deposition rates and further the N saturation threshold still await comprehensive quantization for forest ecosystem. Here, we tested the effect of N addition on aboveground net primary productivity (ANPP) of three-year old Chinese fir (Cunninghamia lanceolata) trees by adding N at 0, 5.6, 11.2, 22.4, and 44.8 g N m-2 yr-1 for 2.5 years. The N saturation threshold was estimated based on a quadratic-plus-plateau model. Results showed that ANPP transitioned from an increasing stage with increasing N addition rate to a plateaued stage at an N rate of 16.3 g N m-2 yr-1. The response of ANPP to N addition rates was well explained by the net photosynthetic rates of needles. Results from the dual isotope measurement [simultaneous determination of needle stable carbon (δ13C) and oxygen (δ18O) isotopes] indicated that the photosynthetic capacity, rather than the stomatal conductance, mediated the response of photosynthesis and ANPP of the young Chinese fir trees to N addition. Accordingly, the amount of needle N partitioning to water-soluble fraction, which is associated with the photosynthetic capacity, also responded to N enrichment with a nonlinear increase. Our study will contribute to a more accurate prediction on the influence of N deposition on C cycles in Chinese fir plantations.

Mesoporous crystalline Ti1-xSnxO2 (0 < x < 1) solid solution for a high-performance photocatalyst under visible light irradiation.

We report a facile and effective inorganic polycondensation combined with aerosol-spray strategy towards high-performance photocatalyst by fabricating mesoporous Ti1-xSnxO2 (0 < x < 1) solid solution. Such Ti1-xSnxO2 nanocrystals with high Sn-doped contents are self-assembled into mesoporous spheres can effectively promote visible-light harvest and high quantum yield, leading a longer lifetime of the photoelectron-hole pairs and less recombination. Such the photocatalysts enhanced photocatalytic activity for the degradation of Rhodamine B (RhB). The representative Ti0.9Sn0.1O2 and Ti0.8Sn0.2O2 compounds reach an optimum degradation of ≈50% and 70%, respectively, after 120 min irradiation under visible irradiation. The mesoporous Ti1-xSnxO2 solid solution could inhibit the recombination of electron-hole pairs, which promote reaction thermodynamics and kinetics for RhB degradation.

Functional identification of PsMYB57 involved in anthocyanin regulation of tree peony.

BACKGROUND: R2R3 myeloblastosis (MYB) genes are widely distributed in plants and comprise one of the largest transcription factor gene families. They play important roles in the regulatory networks controlling development, metabolism, and stress responses. Researches on functional genes in tree peony are still in its infancy. To date, few MYB genes have thus far been reported. RESULTS: In this study, we constructed a comprehensive reference gene set by transcriptome sequencing to obtain R2R3 MYB genes. The transcriptomes of eight different tissues were sequenced, and 92,837 unigenes were obtained with an N50 of 1662 nt. A total of 48,435 unigenes (77.98%) were functionally annotated in public databases. Based on the assembly, we identified 57 R2R3 MYB genes containing full-length open reading frames, which clustered into 35 clades by phylogenetic analysis. PsMYB57 clustered with anthocyanin regulation genes in Arabidopsis and was mainly transcribed in the buds and young leaves. The overexpression of PsMYB57 induced anthocyanin accumulation in tobacco, and four detected anthocyanin structural genes, including NtCHS, NtF3'H, NtDFR, and NtANS, were upregulated. The two endogenous bHLH genes NtAn1a and NtAn1b were also upregulated and may work in combination with PsMYB57 in regulating anthocyanin structural genes. CONCLUSIONS: Our study offers a useful reference to the selection of candidate MYB genes for further functional studies in tree peony. Function analysis of PsMYB57 is helpful to understand the color accumulation in vegetative organs of tree peony. PsMYB57 is also a promising resource to improve plant color in molecular breeding.

Tree peony variegated flowers show a small insertion in the F3'H gene of the acyanic flower parts.

BACKGROUND: The tree peony (Paeonia suffruticosa Andr.) cultivar 'Er Qiao' is appreciated for its unstable variegated flower coloration, with cyanic and acyanic flowers appearing on different branches of the same plant and occasionally in a single flower or petal. However, the variegation mechanism is still unclear. RESULTS: In this study, we found significantly higher contents and more diverse sets of anthocyanins in the cyanic petals than in the acyanic petals. Comparative transcriptome analysis between the two flower types revealed 477 differentially expressed genes (DEGs). Quantitative real-time PCR results verified that the transcript levels of the flavonol synthase (FLS) gene were significantly increased in the acyanic petals. Furthermore, we found that a GCGGCG insertion at 246 bp in the flavonoid 3'-hydroxylase (F3'H) gene-coding region constitutes a duplication of the 241-245 bp section and was consistently found only in acyanic flowers. Sequence alignment of the F3'H gene from different plant species indicated that only the acyanic petals of 'Er Qiao' contained the GCGGCG insertion. The transformation of Arabidopsis tt7-1 lines demonstrated that the ectopic expression of F3'H-cyanic, but not F3'H-acyanic, could complement the colors in the hypocotyl and seed coat. CONCLUSION: In summary, we found that an indel in F3'H and the upregulation of FLS drastically reduced the anthocyanin content in acyanic petals. Our results provide molecular candidates for a better understanding of the variegation mechanisms in tree peony.

The evaluation of the desensitization effect of a desensitizing agent and desensitizing toothpastes in vitro.

This study was evaluating how three desensitizing toothpastes used at home influence the effect associated with desensitizing agents after application in the clinic. Fifty dentine disks measure it permeability and 32 dentine disks with similar permeability levels were selected. Following Dental desensitizer treatment, dentine disks were randomly divided into three subgroups (n=10) that received applications of three toothpastes, respectively. The permeability (Lp) of each specimen was measured after each treatment. One specimen was selected from each group for scanning electron microscopy (SEM) observation. After each treatment, the Lp values decreased significantly for each group (p<0.05) and either completely or partially blocked the dentine tubules upon SEM observation. However, no significant differences in Lp values were observed amongst subgroups (p>0.05). After using the Dental desensitizer, Sensodyne, Crest and Colgate desensitizing toothpastes both can continued to reduce the permeability of the dentine disk, and no significant differences were found amongst them.

The internal interaction in RBBP5 regulates assembly and activity of MLL1 methyltransferase complex.

The Mixed Lineage Leukemia protein 1 (MLL1) plays an essential role in the maintenance of the histone H3 lysine 4 (H3K4) methylation status for gene expression during differentiation and development. The methyltransferase activity of MLL1 is regulated by three conserved core subunits, WDR5, RBBP5 and ASH2L. Here, we determined the structure of human RBBP5 and demonstrated its role in the assembly and regulation of the MLL1 complex. We identified an internal interaction between the WD40 propeller and the C-terminal distal region in RBBP5, which assisted the maintenance of the compact conformation of the MLL1 complex. We also discovered a vertebrate-specific motif in the C-terminal distal region of RBBP5 that contributed to nucleosome recognition and methylation of nucleosomes by the MLL1 complex. Our results provide new insights into functional conservation and evolutionary plasticity of the scaffold protein RBBP5 in the regulation of KMT2-family methyltransferase complexes.

Taxonomic study of Triblemma by using light and scanning electron microscopy.

Triblemma Ching is a genus proposed by Ching Ren-Chang in 1978, and it is composed of two species, Triblemma lancea (Thunb.) Ching and Triblemma zeylanica (Hook.) Ching. There has been much debate on the systematic position of Triblemma, and this genus has always been included in Diplazium. Here, we have described new features of tracheary elements, epidermis, spore, scale, and rachis of Triblemma revealed using light and scanning electron microscopy and proposed that Triblemma is closely related to Athyriopsis, which conflicts with the traditional viewpoint.

Jasmonic acid/ethylene signaling coordinates hydroxycinnamic acid amides biosynthesis through ORA59 transcription factor.

Hydroxycinnamic acid amides (HCAAs) are a class of antimicrobial metabolites involved in plant defense against necrotrophic pathogens, including Alternaria brassicicola and Botrytis cinerea. The agmatine coumaryl transferase (AtACT) is the key enzyme that catalyzes the last reaction in the biosynthesis of HCAAs, including p-coumaroylagmatine (CouAgm) and feruloylagmatine in Arabidopsis thaliana. However, the regulatory mechanism of AtACT gene expression is currently unknown. Yeast one-hybrid screening using the AtACT promoter as bait isolated the key positive regulator ORA59 that is involved in jasmonic acid/ethylene (JA/ET)-mediated plant defense responses. AtACT gene expression and HCAAs biosynthesis were synergistically induced by a combination of JA and ET. In the AtACT promoter, two GCC-boxes function equivalently for trans-activation by ORA59 in Arabidopsis protoplasts, and mutation of either GCC-box abolished AtACT mRNA accumulation in transgenic plants. Site-directed mutation analysis demonstrated that the specific Leu residue at position 228 of the ORA59 EDLL motif mainly contributed to its transcriptional activity on AtACT gene expression. Importantly, MEDIATOR25 (MED25) and ORA59 homodimer are also required for ORA59-dependent activation of the AtACT gene. These results suggest that ORA59 and two functionally equivalent GCC-boxes form the regulatory module together with MED25 that enables AtACT gene expression and HCAAs biosynthesis to respond to simultaneous activation of the JA/ET signaling pathways.

Significant Reduction of the Expression of Peach ( Prunus persica L. Batsch) Allergen-Encoding Genes by Fruit Bagging with Opaque Paper.

Freshly consumed peaches ( Prunus persica L. Batsch) can cause allergic reactions in the worldwide population because of the presence of four classes of allergens (Pru p 1, Pru p 2, Pru p 3, and Pru p 4). Fruit bagging has been widely practiced in peach cultivation to improve fruit quality; however, its effect on the expression of peach allergen-encoding genes remains unknown. In this study, the influence of fruit bagging with opaque paper bags on the major peach allergen-encoding genes, including Pru p 1.01, Pru p 1.06B, Pru p 2.01B, Pru p 2.02, Pru p 3.01, Pru p 4.01, and Pru p 4.02, were measured by means of real-time PCR. A significant reduction in transcript accumulation was observed for all of the selected genes in the epicarps of the bagged peach fruits, whereas slight increases were observed in the mesocarps for these genes, with the two exceptions of Pru p 2.02 and Pru p 3.01. For most of these genes, much higher transcripts were determined in the epicarps than in the mesocarps. Taken together, a significant reduction in the transcription rate of the allergen-encoding genes in the whole peach fruit was achieved by shading with opaque paper bags. According to these data, modifications in growing practices of peach may help to obtain fruits with lower levels of allergens and thus contribute to reducing potential allergenic risks in consumers.

Transcriptome profiling of anthocyanin-related genes reveals effects of light intensity on anthocyanin biosynthesis in red leaf lettuce.

Red leaf lettuce (Lactuca sativa L.) is popular due to its high anthocyanin content, but poor leaf coloring often occurs under low light intensity. In order to reveal the mechanisms of anthocyanins affected by light intensity, we compared the transcriptome of L. sativa L. var. capitata under light intensities of 40 and 100 µmol m-2 s-1. A total of 62,111 unigenes were de novo assembled with an N50 of 1,681 bp, and 48,435 unigenes were functionally annotated in public databases. A total of 3,899 differentially expressed genes (DEGs) were detected, of which 1,377 unigenes were up-regulated and 2,552 unigenes were down-regulated in the high light samples. By Kyoto Encyclopedia of Genes and Genomes enrichment analysis, the DEGs were significantly enriched in 14 pathways. Using gene annotation and phylogenetic analysis, we identified seven anthocyanin structural genes, including CHS, CHI, F3H, F3'H, DFR, ANS, and 3GT, and two anthocyanin transport genes, GST and MATE. In terms of anthocyanin regulatory genes, five MYBs and one bHLH gene were identified. An HY5 gene was discovered, which may respond to light-signaling and regulate anthocyanin structural genes. These genes showed a log2FC of 2.7-9.0 under high irradiance, and were validated using quantitative real-time-PCR. In conclusion, our results indicated transcriptome variance in red leaf lettuce under low and high light intensity, and observed a anthocyanin biosynthesis and regulation pattern. The data should further help to unravel the molecular mechanisms of anthocyanins influenced by light intensity.

Structural basis for activity regulation of MLL family methyltransferases.

The mixed lineage leukaemia (MLL) family of proteins (including MLL1-MLL4, SET1A and SET1B) specifically methylate histone 3 Lys4, and have pivotal roles in the transcriptional regulation of genes involved in haematopoiesis and development. The methyltransferase activity of MLL1, by itself severely compromised, is stimulated by the three conserved factors WDR5, RBBP5 and ASH2L, which are shared by all MLL family complexes. However, the molecular mechanism of how these factors regulate the activity of MLL proteins still remains poorly understood. Here we show that a minimized human RBBP5-ASH2L heterodimer is the structural unit that interacts with and activates all MLL family histone methyltransferases. Our structural, biochemical and computational analyses reveal a two-step activation mechanism of MLL family proteins. These findings provide unprecedented insights into the common theme and functional plasticity in complex assembly and activity regulation of MLL family methyltransferases, and also suggest a universal regulation mechanism for most histone methyltransferases.

Transcriptome sequencing of purple petal spot region in tree peony reveals differentially expressed anthocyanin structural genes.

The pigmented cells in defined region of a petal constitute the petal spots. Petal spots attract pollinators and are found in many angiosperm families. Several cultivars of tree peony contain a single red or purple spot at the base of petal that makes the flower more attractive for the ornamental market. So far, the understanding of the molecular mechanism of spot formation is inadequate. In this study, we sequenced the transcriptome of the purple spot and the white non-spot of tree peony flower. We assembled and annotated 67,892 unigenes. Comparative analyses of the two transcriptomes showed 1,573 differentially expressed genes, among which 933 were up-regulated, and 640 were down-regulated in the purple spot. Subsequently, we examined four anthocyanin structural genes, including PsCHS, PsF3'H, PsDFR, and PsANS, which expressed at a significantly higher level in the purple spot than in the white non-spot. We further validated the digital expression data using quantitative real-time PCR. Our result uncovered transcriptome variance between the spot and non-spot of tree peony flower, and revealed that the co-expression of four anthocyanin structural genes was responsible for spot pigment in tree peony. The data will further help to unravel the genetic mechanism of peony flower spot formation.

The effect of intraoperative fluid volume administration on pancreatic fistulas after pancreaticoduodenectomy.

BACKGROUND: Fluid therapy may be one of the most controversial topics in perioperative management. However, data concerning the influence of perioperative fluid administration on complications after pancreaticoduodenectomy are sparse. METHODS: A group of 147 patients underwent pancreaticoduodenectomy for benign or malignant pathology of the pancreas or the periampullary region between 2005 and 2009. Clinical data, overall morbidity, and long-term outcomes were recorded. RESULTS: We categorized the patients into two groups according to intraoperative fluid volume administration: a low fluid volume group (LFVG, <8.2 ml kg(-1) hr(-1), n = 90) group, and a high fluid volume group (HFVG, ≥8.2 ml kg(-1) hr(-1), n = 57). In terms of colloid administration, the high fluid volume group received significantly more colloid both during the intraoperative period and 0-12 hr after surgery (p < .001 and p < .007, respectively). Pancreatic fistula rates were significantly greater in the high fluid volume group (p = .035). However, the long-term survival rate was not different between the two groups. CONCLUSIONS: High intraoperative fluid volume administration is associated with an increased incidence of pancreatic fistulas after pancreaticoduodenectomy.

Extract of Perilla frutescens inhibits tumor proliferation of HCC via PI3K/AKT signal pathway.

In this study, isoegomaketone(IK) was isolated from Perilla frutescens(L.), a Chinese herbal. The effects of IK were examined by cell viability assay, colony formation assay, xenograft tumor assay and western blotting in HCC cells. We found that IK inhibited cell viability, and its administration decreased tumor volume and weight profoundly. The presence of IK(10 nmol/l) produced a dramatic decrease of pAkt, while total Akt level was not affected. The data suggested that IK from perilla suppressed HCC tumor growth via blocking PI3K/Akt signaling pathway.